Similarity between class A and class B G-protein-coupled receptors exemplified through calcitonin gene-related peptide receptor modelling and mutagenesis studies

Modelling class B G-protein-coupled receptors (GPCRs) using class A GPCR structural templates is difficult due to lack of homology. The plant GPCR, GCR1, has homology to both class A and class B GPCRs. We have used this to generate a class A–class B alignment, and by incorporating maximum lagged correlation of entropy and hydrophobicity into a consensus score, we have been able to align receptor transmembrane regions. We have applied this analysis to generate active and inactive homology models of the class B calcitonin gene-related peptide (CGRP) receptor, and have supported it with site-directed mutagenesis data using 122 CGRP receptor residues and 144 published mutagenesis results on other class B GPCRs. The variation of sequence variability with structure, the analysis of polarity violations, the alignment of group-conserved residues and the mutagenesis results at 27 key positions were particularly informative in distinguishing between the proposed and plausible alternative alignments. Furthermore, we have been able to associate the key molecular features of the class B GPCR signalling machinery with their class A counterparts for the first time. These include the [K/R]KLH motif in intracellular loop 1, [I/L]xxxL and KxxK at the intracellular end of TM5 and TM6, the NPXXY/VAVLY motif on TM7 and small group-conserved residues in TM1, TM2, TM3 and TM7. The equivalent of the class A DRY motif is proposed to involve Arg^2.39, His^2.43 and Glu^3.46, which makes a polar lock with T^6.37. These alignments and models provide useful tools for understanding class B GPCR function.     

Levels of essential and non-essential metals in leaves of the tea plant (Camellia sinensis L.) and soil of Wushwush farms,Ethiopia

Five tea clones of the Camellia assamica variety grown in Wushwush tea plantation farms, Ethiopia, were analyzed for their contents of essential, non-essential and toxic metals (K, Ca, Mg, Fe, Mn, Cu, Zn, Na, Cd and Pb) by atomic absorption flame emission spectroscopy. Both the tea leaves and the soils of the study farms showed similar accumulation patterns in their contents of the studied macronutrients. Among the macronutrient metals, K was the most abundant element in the tea leaves (17.7–24.8 mg/g) and the soils (7.14–9.73 mg/g). Mn was the predominant micronutrient heavy metal in the tea leaf tissues ranging between 501 and 1281 mg/kg. Level of Fe (29.6–100 mg/kg) in the leaf tissue was found to be the second most abundant micronutrient next to Mn whereas concentrations of Cu and Co were relatively lower both in the soil and tea samples. The toxic heavy metals Pb and Cd in the leaf tissues were present at levels too low to be detected by the analytical technique used in this study. The soils were found to be acidic (pH 5.04–5.49) with high organic matter (5.48–6.02%). Fe was the most abundant metal followed by Mn, Na and Zn in the soils. Unlike the tea leaves, the soils were found to contain traces of the toxic metal, Cd (0.02–1.10 mg/kg). The levels of most of the metals determined in this study compared well with those reported for tea leaves from some other parts of the world.     

Current and residual effects of compost and inorganic fertilizer on wheat and soil chemical properties

Restoring soil fertility in smallholder farming systems is essential to sustain crop production. An experiment was conducted in 2011 and 2012 to study the effect of compost and inorganic fertilizer application on soil chemical properties and wheat yield in northwest Ethiopia. Full factorial combinations of four levels of compost (0, 4, 6, 8 t ha^?1) and three levels of inorganic fertilizers (0–0, 17.3–5, 34.5–10 kg N–P ha^?1) were compared in a randomized complete block design with three replications. In 2012, two sets of trials were conducted: one was the repetition of the 2011 experiment on a new experimental plot and the second was a residual effect study conducted on the experimental plots of 2011. Results showed that in the year of application, applying 6 t compost ha^?1 with 34.5–10 kg N–P ha^?1 gave the highest significant grain yield. In the residual effect trial, 8 t compost ha^?1 with 34.5–10 kg N–P ha^?1 gave 271 % increase over the control. Grain protein content increased 21 and 16 % in the current and residual effect trials, respectively, when 8 t compost ha^?1 was applied; it increased 11 and 14 % in the current and residual effect trials, respectively, when 34.5–10 kg N–P ha^?1 was applied. Under the current and residual effects of 8 t compost ha^?1, SOM increased 108 and 104 %; available P 162 and 173 %; exchangeable Ca 16.7 and 17.4 %; and CEC 15.4 and 17.1 %, respectively. Applying 6 t compost ha^?1 with 34.5–10 kg N–P ha^?1 is economically profitable with 844 % MRR.     

The role of ECL2 in CGRP receptor activation: a combined modelling and experimental approach

The calcitonin gene-related peptide (CGRP) receptor is a complex of a calcitonin receptor-like receptor (CLR), which is a family B G-protein-coupled receptor (GPCR) and receptor activity modifying protein 1. The role of the second extracellular loop (ECL2) of CLR in binding CGRP and coupling to Gs was investigated using a combination of mutagenesis and modelling. An alanine scan of residues 271–294 of CLR showed that the ability of CGRP to produce cAMP was impaired by point mutations at 13 residues; most of these also impaired the response to adrenomedullin (AM). These data were used to select probable ECL2-modelled conformations that are involved in agonist binding, allowing the identification of the likely contacts between the peptide and receptor. The implications of the most likely structures for receptor activation are discussed.     

Highly efficient and durable phosphine reduced iron-doped tungsten oxide/reduced graphene oxide nanocomposites for the hydrogen evolution reaction

The design and development of inexpensive and highly efficient electrocatalysts for hydrogen production from water splitting are highly crucial for green energy and the hydrogen economy. Herein, we report phosphine reduced an iron-doped tungsten oxide nanoplate/reduced graphene oxide nanocomposite (Fe-WOxP/rGO) as an excellent electrocatalyst for the hydrogen evolution reaction. This electrocatalyst was synthesized using a hydrothermal method, followed by reduction with phosphine (PH₃), which was generated from sodium hypophosphite. The catalyst onset potential, Tafel slope, and stability were investigated. Accordingly, Fe-WOxP/rGO exhibited impressively high electrocatalytic activity with a low overpotential of 54.60 mV, which is required to achieve a current density of 10 mAcm^?2. The Tafel slope of 41.99 mV dec^?1and the linear sweep voltammetry curve is almost the same as 2000 cycles and electrolysis under static overpotential (54.60 mV) is remain for more than 24 h in 0.5 M H₂SO₄. The catalytic activity and conductivity of Fe-WOxP/rGO were higher than WO_XP, Fe-WOxP and WOxP/rGO. Such an outstanding performance of the Fe-WOxP/rGO nanocomposite is attributed to the coupled synergic effect between high oxygen vacancies formation on tungsten oxide in the nanoplate-like structure of Fe-WOxP and rGO nanosheet, making it as an excellent electrocatalyst for hydrogen evolution reaction.     

Assessment of protein nutritional quality and effects of traditional processes: a comparison between Ethiopian quality protein maize and five Ethiopian adapted normal maize cultivars

The present study was designed to quantitatively measure and compare the levels and variations of total protein, individual amino acids, and computed protein efficiency ratio (C-PER) in raw and traditionally processed products of one recently released quality protein maize (QPM BH542) with four high-yield maize hybrids, namely flint BH660, semi-dent BH140, Pioneer 30H83, and Pioneer 30G97, as well as one local maize cultivar. The total protein content was variable among the cultivars ranging from 7% for BH660 to 8.6% for Pioneer 30H83, 8.9% for BH140, 9.8% for QPM BH542, 10.1% for local maize cultivar, and 11.8% for Pioneer 30G97, respectively. However, the QPM BH542 maize protein proved to be higher in nutritional quality than common maize proteins because it contained 30% to 82% more lysine, higher levels of arginine, tryptophan, histidine, threonine, cysteine, and valine. As a result, the QPM BH542 amino acid profile gives a good balance of total essential amino acids, limited only in lysine, and has a C-PER ratio of 2.2 compared to 1.14, 1.2, 1.4, 1.66, and 1.67 for Pioneer 30G97, local, BH-140, BH660, and Pioneer 30H83, respectively. The various traditional processes of maize have no significant effect on the protein nutritional quality of the new quality protein maize. Hence, the widely dissemination of it in agricultural cultivation as well as consumption by the general population is recommended.